Primitive endoderm stem cell inducer

By employing a Wnt activator or GSK3 inhibitor in a serum-free medium, primitive endoderm stem cells are induced, facilitating the production of artificial blastocyst-like constructs and addressing the lack of established methods for these stem cells.

JP7681903B2Active Publication Date: 2025-05-23THE INSTITUTE OF PHYSICAL & CHEMICAL RESEARCH
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Patent Information

Application Number
JP2021527728
Authority / Receiving Office
JP · JP
Patent Type
Patents
Current Assignee / Owner
Priority Date
2019-06-26
Filing Date
2020-06-25
Publication Date
2025-05-23
Estimated Expiration
2040-06-25

AI Technical Summary

Technical Problem

There is no known method for establishing primitive endoderm stem cells, which are essential for artificially reconstructing a blastocyst by combining stem cells of different types.

Method used

The use of a Wnt activator or a GSK3 inhibitor, such as CHIR99021, in a serum-free medium to induce primitive endoderm stem cells, allowing for the production of artificial blastocyst-like constructs.

Benefits of technology

This method effectively induces primitive endoderm stem cells and enables the creation of artificial blastocyst-like constructs, which can be used to determine their implantation competence.

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Abstract

The present invention pertains to: a primitive endoderm stem cell inducer containing a Wnt activator or a GSK3 inhibitor; a medium which is for inducing a primitive endoderm stem cell, contains said inducer, and does not contain serum; and a method which is for producing a primitive endoderm stem cell and comprises culturing a blastocyst in a serum-free medium containing a Wnt activator or a GSK3 inhibitor.
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Claims

1. A primitive endoderm stem cell inducer, comprising CHIR99021.

2. A medium for primitive endoderm stem cell induction comprising CHIR99021 and free of serum.

3. The medium according to claim 2, further comprising no PD0325901.

4. 1. A method for producing primitive endoderm stem cells comprising culturing blastocysts in a serum-free medium containing CHIR99021.

5. 5. The method of claim 4, wherein the medium does not contain PD0325901.

6. The method according to claim 4 or 5, wherein the cells are excluding cells of human origin.

7. 1. A method for producing a blastocyst-like construct, comprising mixing a blastocyst with primitive endoderm stem cells, epiblast stem cells and / or ES cells, and trophoblast stem cells, obtained by culturing the blastocyst in a serum-free medium containing CHIR99021, All stem cells, except those derived from humans, Manufacturing method.

8. The method according to claim 7, wherein the medium does not contain PD0325901.

9. The method according to claim 7 or 8, wherein the cells are not cells derived from humans.

10. Use of CHIR99021 in inducing primitive endoderm stem cells.

11. Use of serum-free medium containing CHIR99021 in the induction of primitive endoderm stem cells.

12. 12. The use according to claim 10 or 11, wherein the cells are excluding cells of human origin.

13. Mixing the blastocyst with primitive endoderm stem cells, epiblast stem cells and / or ES cells obtained by culturing in a serum-free medium containing CHIR99021, and trophoblast stem cells to obtain a blastocyst-like construct; and implanting the resulting blastocyst-like construct into a non-human uterus.

1. A method for determining the implantation competence of an implanted blastocyst-like construct, comprising: All stem cells, except those derived from humans, method.

Citation Information

Patent Citations

  • Culture medium containing kinase inhibitor, and use thereof

    JP2013066495A

  • Trophoblast ectoderm-like structure, and method for producing the same

    JP2016214138A

  • Primitive endodermal stem cells, a process for preparing them and their use, in particular for obtaining primitive epithelial liver cells

    WO2006128681A1

  • Method for establishing and maintaining trophoblast stem cells

    WO2015125926A1

  • Method for inducing primitive endoderm from naive pluripotent stem cells

    WO2019093340A1